Complex microbial inoculant, preparation method thereof and application of complex microbial inoculant in promoting compost fermentation
By using a compound inoculant of Bacillus licheniformis and Bacillus coagulans in cow manure composting, the problem of slow temperature rise during cow manure fermentation in winter was solved, achieving rapid temperature rise and shortening the fermentation cycle, thus improving composting quality and efficiency.
Patent Information
- Application Number
- CN202511558368.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-10-29
- Publication Date
- 2026-01-27
AI Technical Summary
Traditional aerobic fermentation of cow manure has problems such as difficulty in raising the fermentation temperature in winter, long fermentation cycle, and uneven quality. In particular, the temperature rises slowly in the fermentation of cow manure in winter, and the temperature in the core area of the stack is low.
A thermophilic and heat-resistant compound microbial agent, including Bacillus licheniformis and Bacillus coagulans, is inoculated, mixed in a specific ratio, and combined with a substrate (such as corn cobs) to prepare a compound microbial agent for use in cow manure composting fermentation to improve the heating rate and core temperature.
It significantly shortens the winter fermentation start-up time by 20% to 70%, increases the core temperature by 10 to 50°C, shortens the fermentation cycle from 12 to 16 days to 5 to 6 days, reduces the moisture content by 10% to 40%, and improves fermentation efficiency and sterilization effect.
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Abstract
Description
Technical Field
[0001] This invention relates to the field of microbial fermentation technology, and more specifically, to a compound microbial agent, its preparation method, and its application in promoting compost fermentation. Background Technology
[0002] In recent years, with the rapid development of domestic animal husbandry and the continuous increase in the number of modern ranches, cow manure treatment has become an important environmental challenge for the domestic breeding industry. Fermenting cow manure into cow bedding material is one of the important means to achieve green recycling of breeding waste. However, traditional aerobic fermentation has problems such as difficulty in raising the fermentation temperature in winter, long fermentation cycle, and uneven quality. Therefore, there is an urgent need for a fermentation agent that can quickly promote the temperature rise of compost and shorten the composting fermentation cycle. Summary of the Invention
[0003] This invention addresses the problems of slow temperature rise and low temperature in the core area of cow manure fermentation during winter. It employs inoculation of the stack with a thermophilic and heat-resistant compound microbial agent to increase the temperature rise rate and the core area temperature, thereby achieving both fermentation and sterilization effects.
[0004] To achieve the above objectives, the present invention is implemented through the following technical solution: In a first aspect, the present invention provides a compound bacterial agent comprising a compound bacterial solution and a matrix, wherein the compound bacterial solution contains Bacillus licheniformis and Bacillus coagulans; the mass ratio of Bacillus licheniformis to Bacillus coagulans is 1:(0.4~2.5).
[0005] In a preferred embodiment, the mass ratio of Bacillus licheniformis to Bacillus coagulans is 1:1.5.
[0006] In the preferred embodiment, the total number of viable bacteria in the compound bacterial solution is 10. 7 ~10 9 CFU / g; more preferably, the total number of viable bacteria in the compound bacterial solution is 10. 8 CFU / g.
[0007] In a preferred embodiment, the substrate is selected from one or more of corn cobs, cotton stalks, rice husks, and wheat bran.
[0008] In a preferred embodiment, the mass ratio of the substrate to the composite bacterial solution is (0.1~0.5):1.
[0009] In a preferred embodiment, the mass ratio of the substrate to the composite bacterial solution is 0.33:1.
[0010] A second aspect of the present invention provides a method for preparing the compound microbial agent described in any of the above embodiments, comprising the following steps: S1. After activating and culturing Bacillus licheniformis and Bacillus coagulans respectively, they were combined at a mass ratio of 1:(0.4~2.5), resuspended, and a compound bacterial solution was obtained. S2. Add the compound bacterial solution to the substrate and air dry it naturally to obtain the compound bacterial agent.
[0011] A third aspect of the present invention provides the application of the compound microbial agent described in any of the above embodiments in promoting compost fermentation.
[0012] Under preferred embodiments, the application of the above-mentioned compound microbial agent in promoting compost fermentation includes any one or more of the following: 1) Shorten the fermentation cycle of cow manure; 2) Promotes composting temperature rise.
[0013] In the preferred embodiment, the above-mentioned compound microbial agent is applied to promote composting fermentation by adding 100-300g of compound microbial agent per ton of cow manure; more preferably, 200g of compound microbial agent is added per ton of cow manure.
[0014] Beneficial effects of the present invention The compound microbial agent provided by this invention can shorten the start-up time of winter fermentation by 20% to 70%, increase the temperature of the core area by 10 to 50°C, shorten the fermentation cycle from the conventional 12 to 16 days to 5 to 6 days, and reduce the moisture content by 10% to 40%. Attached Figure Description
[0015] Figure 1 This describes the temperature changes on day 1 of composting fermentation in Examples 1-7 and Comparative Examples 1-4 of the present invention. Figure 2 This describes the temperature changes during composting fermentation of Examples 1-7 and Comparative Examples 1-4 of the present invention from day 1 to day 16. Figure 3 The moisture content after composting fermentation in Examples 1-7 and Comparative Examples 1-4 of this invention is measured. Detailed Implementation
[0016] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of the present invention.
[0017] Unless otherwise stated, scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. Unless otherwise specified, the experimental methods used in the examples are conventional methods. Where specific conditions are not specified in the examples, they should be performed under conventional conditions or conditions recommended by the manufacturer. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.
[0018] 1. Test strain The *Bacillus licheniformis* and *Bacillus coagulans* used in this invention were purchased from the China General Microbiological Culture Collection Center (CGMCC). *Bacillus licheniformis* ( Bacillus licheniformis BGB-84F, accession number CGMCC No. 25764; Bacillus coagulans ( Bacillus coagulans ) BC2000, accession number CGMCCNo.21621.
[0019] 2. Test culture medium LB liquid medium: tryptone 10 g / L, yeast extract 5 g / L, NaCl 10 g / L, diluted with distilled water to 1 L; LB solid medium: obtained by adding 10 g / L agar powder to the above LB liquid medium; Beef broth culture medium: 10 g / L protein, 15 g / L beef extract, 15 g / L NaCl, diluted to 1 L with distilled water.
[0020] 3. Culture of the tested strains Bacillus licheniformis and Bacillus coagulans were activated and cultured according to the following steps, the specific process of which is as follows: 1) Activation culture: The bacterial strain was inoculated into LB solid medium at an inoculation rate of 5% and cultured at 30°C for 48 hours to obtain the activated bacterial strain; 2) Fermentation culture: The activated strain was inoculated into beef broth medium at an inoculation rate of 3%, and cultured on a shaker at 37°C for 12 hours at a speed of 150 rpm. The medium was then discarded to obtain the fermented strain.
[0021] Example 1
[0022] A method for preparing a compound microbial agent includes the following steps: S1. Mix Bacillus licheniformis and Bacillus coagulans, which have been activated and fermented, at a mass ratio of 7:3, and resuspend them in LB liquid medium until the total number of viable bacteria is 10. 8 CFU / g was used to obtain a compound bacterial solution; S2. Crush the corn cob, weigh 66g of the crushed corn cob, add 200g of compound bacterial solution, and air dry naturally to obtain the compound bacterial agent.
[0023] Example 2
[0024] A method for preparing a compound microbial agent includes the following steps: S1. Mix Bacillus licheniformis and Bacillus coagulans, which have been activated and fermented, at a mass ratio of 3:2, and resuspend them in LB liquid medium until the total number of viable bacteria is 10. 8 CFU / g was used to obtain a compound bacterial solution; S2. Crush the corn cob, weigh 66g of the crushed corn cob, add 200g of compound bacterial solution, and air dry naturally to obtain the compound bacterial agent.
[0025] Example 3
[0026] A method for preparing a compound microbial agent includes the following steps: S1. Mix Bacillus licheniformis and Bacillus coagulans, which have been activated and fermented, at a mass ratio of 1:1, and resuspend them in LB liquid medium until the total number of viable bacteria is 10. 8 CFU / g was used to obtain a compound bacterial solution; S2. Crush the corn cob, weigh 66g of the crushed corn cob, add 200g of compound bacterial solution, and air dry naturally to obtain the compound bacterial agent.
[0027] Example 4
[0028] A method for preparing a compound microbial agent includes the following steps: S1. Mix Bacillus licheniformis and Bacillus coagulans, which have been activated and fermented, at a mass ratio of 2:3, and resuspend them in LB liquid medium until the total number of viable bacteria is 10. 8 CFU / g was used to obtain a compound bacterial solution; S2. Crush the corn cob, weigh 66g of the crushed corn cob, add 200g of compound bacterial solution, and air dry naturally to obtain the compound bacterial agent.
[0029] Example 5
[0030] A method for preparing a compound microbial agent includes the following steps: S1. Mix Bacillus licheniformis and Bacillus coagulans, which have been activated and fermented, at a mass ratio of 3:7, and resuspend them in LB liquid medium until the total number of viable bacteria is 10. 8 CFU / g was used to obtain a compound bacterial solution; S2. Crush the corn cob, weigh 66g of the crushed corn cob, add 200g of compound bacterial solution, and air dry naturally to obtain the compound bacterial agent.
[0031] Example 6
[0032] A method for preparing a compound microbial agent includes the following steps: S1. Mix Bacillus licheniformis and Bacillus coagulans, which have been activated and fermented, at a mass ratio of 2:3, and resuspend them in LB liquid medium until the total number of viable bacteria is 10.8 CFU / g was used to obtain a compound bacterial solution; S2. Crush the corn cob, weigh 20g of the crushed corn cob, add 200g of compound bacterial solution, and air dry naturally to obtain the compound bacterial agent.
[0033] Example 7
[0034] A method for preparing a compound microbial agent includes the following steps: S1. Mix Bacillus licheniformis and Bacillus coagulans, which have been activated and fermented, at a mass ratio of 2:3, and resuspend them in LB liquid medium until the total number of viable bacteria is 10. 8 CFU / g was used to obtain a compound bacterial solution; S2. Crush the corn cob, weigh 100g of the crushed corn cob, add 200g of compound bacterial solution, and air dry naturally to obtain the compound bacterial agent.
[0035] Comparative Example 1 A method for preparing a compound microbial agent includes the following steps: S1. The activated and fermented Bacillus licheniformis was resuspended in LB liquid medium until the total number of viable bacteria was 10. 8 CFU / g was used to obtain a compound bacterial solution; S2. Crush the corn cob, weigh 66g of the crushed corn cob, add 200g of compound bacterial solution, and air dry naturally to obtain the compound bacterial agent.
[0036] Comparative Example 2 A method for preparing a compound microbial agent includes the following steps: S1. The activated and fermented Bacillus coagulans were resuspended in LB liquid medium until the total number of viable bacteria was 10. 8 CFU / g was used to obtain a compound bacterial solution; S2. Crush the corn cob, weigh 66g of the crushed corn cob, add 200g of compound bacterial solution, and air dry naturally to obtain the compound bacterial agent.
[0037] Comparative Example 3 A method for preparing a compound microbial agent includes the following steps: S1. Mix Bacillus licheniformis and Bacillus coagulans, which have been activated and fermented, at a mass ratio of 10:1, and resuspend them in LB liquid medium until the total number of viable bacteria is 10. 8 CFU / g was used to obtain a compound bacterial solution; S2. Crush the corn cob, weigh 100g of the crushed corn cob, add 200g of compound bacterial solution, and air dry naturally to obtain the compound bacterial agent.
[0038] Comparative Example 4 A method for preparing a compound microbial agent includes the following steps: S1. Mix Bacillus licheniformis and Bacillus coagulans, which have been activated and fermented, at a mass ratio of 1:10, and resuspend them in LB liquid medium until the total number of viable bacteria is 10. 8 CFU / g was used to obtain a compound bacterial solution; S2. Crush the corn cob, weigh 100g of the crushed corn cob, add 200g of compound bacterial solution, and air dry naturally to obtain the compound bacterial agent.
[0039] Test on the effect of compound microbial agents on compost fermentation: 1. Experimental Grouping The compost was divided into 14 groups according to the different compound microbial agents added. The 14 groups of compost were respectively added with the compound microbial agents prepared in Examples 1 to 7 and Comparative Examples 1 to 4.
[0040] 2. Test cow dung Cow dung: provided by Tianrun Fenghuotai Ranch.
[0041] 3. Experimental Methods Cow manure from the pasture is scraped onto a manure storage pit, then passes through a dry-wet separation room and undergoes a primary dry-wet separation system to obtain cow manure with a moisture content of 65%–70%. After natural sun drying, the moisture content is reduced to approximately 60%, at which point it is ready for composting. Different compound microbial agents are added to the cow manure of each experimental group at a dosage of 200g / ton, with an initial ambient temperature of -20℃. The mixed materials are thoroughly stirred and piled into windrows 2m wide and 1m high, with ventilation openings at the bottom. Ventilation is carried out for 2 hours daily for one week. After one week, the ventilation frequency is reduced to 1 hour every 2 days, with the pile turned over every 3 days to continue fermentation. The fermentation cycle for each group of compost is determined based on the fermentation progress. After fermentation is complete, the compost is cooled to ambient temperature.
[0042] 4. Compost Index Testing Methods 4.1 Compost Fermentation Cycle Test Sampling was conducted once every day, with five sampling points taken at heights of 30cm, 50cm, and 70cm in the compost. After mixing, cucumber seeds were cultured using each group of compost according to industry standard NY / T 525-2021, and the seed germination index of the cucumber seeds was tested. The compost fermentation period was determined with a seed germination index ≥90%. Fermentation was terminated when the seed germination index ≥90%, and the number of days of compost fermentation was recorded.
[0043] 4.2 Composting Temperature Test The temperature of the compost was measured using a penetration thermometer at heights of 30cm, 50cm, and 70cm. The test points are as follows: Figure 1As shown, on the first day of composting fermentation, the temperature was measured every 4 hours. Thereafter, the temperature was measured every 24 hours at 10:00 AM. The average value of the temperature measured in each time period was taken and retained to one decimal place.
[0044] 4.3 Fertilizer Moisture Content Test Five-point sampling was used to collect samples at heights of 30cm, 50cm, and 70cm in the compost. After mixing, the moisture content of each compost group after fermentation was measured using a halogen moisture analyzer.
[0045] 5. Test Results 5.1 Composting Fermentation Cycle The fermentation cycle of compost treated by each catalyst group is shown in Table 1. The results show that, compared with the composite microbial agents prepared in Comparative Examples 1-4, the compost fermentation cycle can be significantly shortened. Among them, the composite microbial agent prepared in Example 4 has the shortest fermentation cycle, which can complete the compost fermentation in only 5 days.
[0046] Table 1 Composting Fermentation Cycle
[0047] 5.2 Fertilizer Temperature Test Results The temperature changes of each group on day 1 of composting fermentation are as follows: Figure 1 As shown in the figure. The results indicate that, compared to the composite microbial agents prepared in Comparative Examples 1-4, the composite microbial agents prepared in Examples 1-7 can significantly shorten the start-up time of compost fermentation. After 1 day of fermentation, the compost temperature treated with the composite microbial agents prepared in Examples 1-7 for 1 day can all rise to above 50℃. The temperature changes of each group of compost fermentation from day 1 to day 16 are shown in the figure. Figure 2 As shown in the figure. The results indicate that the fermentation temperature of compost treated with the compound microbial agents prepared in Examples 1-7 was significantly higher than that of compost treated with the compound microbial agents prepared in Comparative Examples 1-4.
[0048] Therefore, mixing Bacillus licheniformis and Bacillus coagulans in a specific ratio can significantly increase the compost fermentation temperature and shorten the fermentation start-up time and cycle. The compound microbial agent (2:3) prepared in Example 4 showed the best temperature-raising effect, with the compost temperature rising to 77.6℃ after 1 day of treatment, and remaining stable at around 70℃ during fermentation. Comparing the compost temperature measurements after treatment with the compound microbial agents prepared in Examples 4 and 6-7, it was found that the matrix content in the compound microbial agent is also one of the factors affecting the compost temperature rise. The fermentation temperature showed a trend of first increasing and then decreasing with the increase of matrix content, and the effect was best when the mass ratio of matrix to compound microbial liquid in the compound microbial agent was 0.33:1.
[0049] 5.3 Results of fertilizer moisture content test The changes in moisture content of each compost group after fermentation are as follows: Figure 3As shown. The results indicate that, compared with the compost treated with the composite microbial agents prepared in Comparative Examples 1-4, the compost treated with the composite microbial agents prepared in Examples 1-7 can significantly reduce the moisture content of the compost, saving subsequent dehydration and drying processes, and effectively reducing the cost of compost delivery.
[0050] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.
Claims
1. A compound microbial agent, characterized in that, It includes a compound bacterial solution and a matrix, wherein the compound bacterial solution contains Bacillus licheniformis and Bacillus coagulans; the mass ratio of Bacillus licheniformis to Bacillus coagulans is 1:(0.4~2.5).
2. The compound microbial agent according to claim 1, characterized in that, The mass ratio of Bacillus licheniformis to Bacillus coagulans is 1:1.
5.
3. The compound microbial agent according to claim 1, characterized in that, The total number of live bacteria in the compound bacterial solution is 10. 7 ~10 9 CFU / g.
4. The compound microbial agent according to claim 1, characterized in that, The substrate is selected from one or more of corn cobs, cotton stalks, rice husks, and wheat bran.
5. The compound microbial agent according to claim 1, characterized in that, The mass ratio of the substrate to the composite bacterial solution is (0.1~0.5):
1.
6. The compound microbial agent according to claim 5, characterized in that, The mass ratio of the substrate to the composite bacterial solution is 0.33:
1.
7. A method for preparing the compound microbial agent according to any one of claims 1 to 6, characterized in that, Includes the following steps: S1. After activating and culturing Bacillus licheniformis and Bacillus coagulans respectively, they were combined at a mass ratio of 1:(0.4~2.5), resuspended, and a compound bacterial solution was obtained. S2. Add the compound bacterial solution to the substrate and air dry it naturally to obtain the compound bacterial agent.
8. The application of the compound microbial agent according to any one of claims 1 to 6 in promoting compost fermentation.
9. The application of the compound microbial agent according to claim 8 in promoting compost fermentation, characterized in that, Includes one or more of the following: 1) Shorten the fermentation cycle of cow manure; 2) Promotes composting temperature rise.
10. The application of the compound microbial agent according to claim 8 in promoting compost fermentation, characterized in that, Add 100-300g of compound microbial agent per ton of cow manure.